Figure 5. Poliovirus infection does not block HIV-1 pr55^Gag synthesis but dramatically reduces 4kb- and 2kb-encoded viral gene products, Vpr and Tat.

(A). Schematic representation of the experimental protocol. (B) Whole cell lysates were analyzed for viral pr55^Gag and gp160/gp120 (Env) and host proteins eIF4GI, eIFGII and PABP by western blotting analyses. GAPDH served as a loading control (bottom). (C,D) At the time points indicated post-infection, cells were briefly pulsed with radiolabelled amino acids and immediately harvested for immunoprecipitation analysis. Metabolically-labeled proteins found in whole cell lysates (Panel C (cells), Panel D (left panel (cells)) and cell-free virus lysates (Panel D (right panel (virus)) were immunoprecipitated using human anti-HIV-1 No. 162. Cell lysates were sequentially immunoprecipitated with rabbit anti-Tat and anti-Vpr antisera (Panel D, left panel) and revealed by fluorography. Fractionation was performed by SDS-PAGE (either 10 or 14% polyacrylamide gels) as described in Materials and Methods. Non-specific bands are indicated (NS) and the poliovirus Capsid proteins (2BC, Vp0, Vp1, Vp2 & Vp3) and low molecular weight (*) poliovirus proteins found in immunoprecipitates are indicated; molecular weights are indicated on left.